辽宁省葫芦岛地区鼠类汉坦病毒的分离与鉴定  

Isolation and characterization of Hantavirus carried by rodents in Huludao, Liaoning province

作　　者：屈勇刚[1] 杨国庆[2] 邹洋 剡根强[1] 陈化新[3] 张永振[3] 

机构地区：[1]新疆石河子大学动物科技学院,石河子832003 [2]辽宁省葫芦岛市疾病预防控制中心 [3]中国疾病预防控制中心传染病预防控制所

出　　处：《中华流行病学杂志》2006年第6期513-517,共5页Chinese Journal of Epidemiology

基　　金：科技部科研院所社会公益研究专项资金项目(2002DIB40095);国家"十五"科技攻关课题资助项目(2003BA712A08-02)

摘　　要：目的研究葫芦岛地区鼠类中汉坦病毒的感染流行情况以及病毒的型别。方法采用夹夜法捕捉鼠类,间接免疫荧光法检测鼠肺中的汉坦病毒抗原,阳性较强的标本接种到Vero E6细胞分离病毒,RT-PCR对分离的病毒与阳性样品扩增核苷酸片段并测序,构建系统发生树进行分型与系统发生分析。结果在200份鼠肺标本中共检测到11个样品阳性,阳性率为5．5％。选择阳性较强的标本,接种到Vero E6细胞并连续传代后分离到3株病毒。用S片段(620-999 nt)与M片段G1区(180-580 nt)的核苷酸构建的系统发生树,结果表明葫芦岛分离的3株病毒均为S3亚型。用G2区的2003-2302 nt的核苷酸序列构建的系统发生树将来自不同地区的SEO型病毒可分为7个亚型,其中从葫芦岛地区分离的3株病毒与北京地区分离株CP211、ch302、dc501和山东省分离株SD10、SD227株的亲缘关系最近。结论葫芦岛地区鼠类中汉坦病毒的携带率较高,主要流行S3亚型SEOV。Objective To investigate the Hantavirus infection and their genotype in rodents in Huludao. Methods Rodents were collected from the main epidemic areas to detect antigen of Hantavirus in rat lungs by indirect immunofluorescence assay. Antlgen-positlve samples were inoculated onto cultures of confluent Veto E6 cells for the isolation of virus. The genotypes of viruses in all antigen-positive samples were identified by reverse transcriptase-polymerase chain reaction （RT-PCR）. Results 200 rats were collected in the main epidemic areas, and 11 Hantavirus-positive samples were tested. The positive rate of Hantavirus in rats was 5.5 %, Three strains of Hantavirus were isolated in Veto E6 cell culture. Data from the phylogenetic trees constructed by partial S segment （620-999 nt） or partial G1 segment （180-580 nt） showed that the three isolates carried by rats from Huludao were all genetic subtype SEOV 3. Furthermore, the phylogenetic tree constructed by partial G2 segment （2003-2302 nt） divided SEOV strains into 7 genetic subtypes, and the three isolates were having a closer evolutionary relationship with isolates CP211, ch302 and dc501 from Beijing, and the isolates SD10 and SD227 form Shandong. Conclusion Data indicated that the rate of carrying virus was high and the main genetic subtype of Hantavirus was S3 of Seoul virus in Huludao area.

关 键 词：汉坦病毒 病毒分离 逆转录-聚合酶链反应 系统发生分析 

分 类 号：S852.65[农业科学—基础兽医学]